US20210230525A1 - Oocyte-holding pipette for sperm injection methods without cytoplasmic aspiration - Google Patents

Oocyte-holding pipette for sperm injection methods without cytoplasmic aspiration Download PDF

Info

Publication number
US20210230525A1
US20210230525A1 US17/052,306 US201917052306A US2021230525A1 US 20210230525 A1 US20210230525 A1 US 20210230525A1 US 201917052306 A US201917052306 A US 201917052306A US 2021230525 A1 US2021230525 A1 US 2021230525A1
Authority
US
United States
Prior art keywords
oocyte
microns
diameter
funnel
sperm injection
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US17/052,306
Other languages
English (en)
Inventor
Francisco VERGARA ALCAIDE
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Publication of US20210230525A1 publication Critical patent/US20210230525A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/06—Bioreactors or fermenters specially adapted for specific uses for in vitro fertilization
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
    • A61D19/00—Instruments or methods for reproduction or fertilisation
    • A61D19/04—Instruments or methods for reproduction or fertilisation for embryo transplantation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B17/00—Surgical instruments, devices or methods, e.g. tourniquets
    • A61B17/42—Gynaecological or obstetrical instruments or methods
    • A61B17/425—Gynaecological or obstetrical instruments or methods for reproduction or fertilisation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B17/00—Surgical instruments, devices or methods, e.g. tourniquets
    • A61B17/42—Gynaecological or obstetrical instruments or methods
    • A61B17/425—Gynaecological or obstetrical instruments or methods for reproduction or fertilisation
    • A61B17/43—Gynaecological or obstetrical instruments or methods for reproduction or fertilisation for artificial insemination
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/04—Cell isolation or sorting

Definitions

  • the invention relates to an oocyte-holding pipette for sperm injection methods without cytoplasmic aspiration, providing the intended function thereof with advantages and novelty features that are described in detail below and constitute an alternative improvement in the current state of the art.
  • the object of the invention focuses on a micropipette for use as means for holding an oocyte in intracytoplasmic sperm injection or ICSI methods, the novel structural configuration of which, unlike conventional pipettes, eschews the step of aspirating the cytoplasm with the injection needle to ensure the rupture of the cytoplasmic membrane, since the new pipette, provided with a flared end, holds the oocyte by suction, slightly deforming it inside said end, and said deformation causes an increase in oocyte turgor which allows breaking the cytoplasmic membrane with the injection pipette without first suctioning the cytoplasm.
  • the field of application of the present invention is comprised within the sector of the industry dedicated to the manufacture of medical instruments, focusing particularly on the field of microdevices applicable for cellular-level procedures and interventions, and more specifically to devices intended for use in assisted reproduction procedures.
  • intracytoplasmic sperm injection or ICSI is an assisted reproduction technique consisting of the fertilization of oocytes by means of injecting a sperm into their cytoplasm using a micropipette, after obtaining and preparing the gametes for the purpose of obtaining embryos that can be transferred to the mother's womb.
  • the previously denuded oocyte must be held with a holding pipette, while the sperm is injected with another pipette, in this case an injection pipette.
  • the problem is that the membrane surrounding the oocyte is a hard-to-break membrane due to its high elasticity; in fact, the injection needle can be inserted into the cytoplasm without actually breaking the membrane (the membrane was microscopically invaginated by the injection pipette but did not break).
  • the conventional way for breaking the membrane is to perform aspiration with the injection pipette until breaking the membrane.
  • this leads to a disorganized cytoplasm that may critically affect the mitotic spindle (the location of which could only be estimated as it is not observed with routine ICSI technique) if its arrangement is altered during cytoplasmic aspiration.
  • the membrane must break for the proper release of the sperm into the cytoplasm, and also for the activation of the Ca++ or calcium ion channel mechanisms, which in turn intervene in the activation of cell division mechanisms.
  • the applicant himself is the proprietor of utility model ES1187335U which describes a “pipette for retaining oocytes in intracytoplasmic sperm injection methods” the features of which allow holding the oocyte and performing injection through its membrane without prior cytoplasmic aspiration.
  • the configuration of said holding pipette has an innovative working end holding the oocyte by means of pressure, for which it has a decreasing frustoconical portion where the diameter of its cylindrical body is significantly reduced until defining a small support tip with a planar, straight, and angled configuration with respect to the cylindrical body.
  • Said holding pipette therefore holds the oocyte by means of a slight pressure, not by means of suction like conventional pipettes, which pressure causes an increase in oocyte turgor and thereby allows breaking the cytoplasmic membrane without having to aspirate the cytoplasm.
  • the objective of the present invention is to develop another different, alternative holding pipette model that does not require aspirating the cytoplasm with the injection pipette either, which model, unlike the one mentioned above, provides greater assurance when holding the oocyte, as this is an aspect of the pipette susceptible to improvement.
  • the present invention acts by means of suctioning the oocyte with the holding pipette itself, just like conventional pipettes with the only difference being that while those (conventional) pipettes only perform aspiration to hold the oocyte without deforming it, this pipette (the pipette of the present invention) aspirates the oocyte to deform it while it is partially housed in the flared working end thereof, as will be explained in detail below.
  • the oocyte-holding pipette for sperm injection methods without cytoplasmic aspiration proposed by the invention is therefore configured as a remarkable novelty in its field of application, given that the objectives indicated above are specifically and satisfactorily achieved in view of its implementation, with the characterizing details making it possible and distinguishing same being suitably described in the final claims accompanying the present description.
  • the invention proposes a pipette for holding oocytes in intracytoplasmic sperm injection methods which fulfills said purpose of holding without cytoplasmic aspiration, as it eschews such step of performing aspiration with the injection needle, which is required in conventional holding pipettes to ensure the rupture of the cytoplasmic membrane and cause the activation of the oocyte and to more readily deposit the sperm in the cytoplasm, but which, as pointed out in the preceding section, may damage the original structure of the cytoplasm and, in some cases, alter the structure of the mitotic spindle.
  • the holding pipette of the present invention comprises an elongate and narrow cylindrical body, the working end of which, i.e., the end used for holding the oocyte, is characterized by having a flared funnel-shaped configuration communicating through its smaller-diameter end with the aspiration conduit running along the cylindrical body, the larger opposite end being sized to receive the oocyte in a tight-fitting manner, such that, upon aspirating said oocyte, it remains trapped inside said flared end and becomes deformed enough so as to have a more turgid surface which facilitates the application of the injection pipette without having to previously aspirate the cytoplasm with the pipette.
  • the pipette without cytoplasmic aspiration of the invention fulfills the purpose of holding the oocyte in ICSI methods but eschews the step of aspirating the cytoplasm with the injection needle, which is required in conventional holding pipettes having a rounded working end to ensure the rupture of the cytoplasmic membrane and cause the activation of the oocyte and to more readily deposit the sperm in the cytoplasm, but which damages the original structure of the cytoplasm and, in some cases, may alter the structure of the mitotic spindle.
  • the holding pipette of the invention is manufactured from embryo-tested glass, specifically borosilicate, and the method for manufacturing the holding pipette of the invention is also similar to the method for manufacturing these conventional holding pipettes.
  • the dimensions of the pipette and its parts are conceived so that it can be used in the same microinjection equipment in which conventional holding pipettes are used, only slightly changing the ICSI technique protocol.
  • the described pipette therefore represents an innovation in structural and constitutive features that have been unknown up until now, and these reasons, combined with its practical usefulness, provide it with sufficient grounds to obtain the exclusive privilege that is sought.
  • FIG. 1 shows a side elevational view of the holding pipette object of the invention, with its general configuration and the main parts it comprises being observed.
  • FIG. 2 shows an enlarged view of detail A indicated in FIG. 1 and corresponding to the working end of the pipette, according to the invention, with the general configuration thereof being observed.
  • FIGS. 3 and 4 show respective enlarged views according to a longitudinal section view and an enlarged front elevational view, respectively, of the working end of the pipette shown in FIG. 2 , with the configuration and dimensions thereof being observed in greater detail.
  • FIGS. 5 and 6 show respective schematic views of the mode of operation of the working end of the pipette of the invention for holding the oocyte by means of suction and deformation of the oocyte and without aspiration of its cytoplasm.
  • the holding pipette ( 1 ) of the invention comprises an elongate, narrow, and hollow cylindrical body ( 100 ), inside which runs a suction conduit ( 101 ) that communicates with a working end ( 1 a ) for holding the oocyte ( 2 ), which end is characterized by having a flared funnel-shaped configuration defining an internal conical cavity ( 105 ), the smaller-diameter proximal end ( 102 ) of which communicates with the mentioned suction conduit ( 101 ), the larger-diameter distal end ( 103 ) being sized to receive the oocyte ( 2 ) in a tight-fitting manner, such that, upon aspirating same via suction conduit ( 101 ), the oocyte remains partially trapped inside and becomes deformed, as shown in FIGS.
  • the cylindrical body ( 100 ) has a length (L 1 ) of about 5 cm and an external diameter (D) of 500 micra
  • the suction conduit ( 101 ) has a constant internal diameter (d) of 80 micra along its entire length with the exception of the point at which it is joined to the funnel-shaped working end ( 1 a ), there being, in the outlet opening of said conduit ( 101 ), coinciding with the smaller-diameter proximal end ( 102 ) of said funnel, a constriction ( 104 ) where said diameter is reduced considerably to only about 17 micra.
  • the internal conical cavity ( 105 ) defining said funnel-shaped working end ( 1 a ) also preferably has a length (L 2 ) of 128 micra and an internal diameter (d), at its larger-diameter distal end ( 103 ), of 120 micra, whereas the external diameter (D) of said distal end ( 103 ) of the working end ( 1 a ) is preferably 180 micra.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Surgery (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Reproductive Health (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pregnancy & Childbirth (AREA)
  • Gynecology & Obstetrics (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Medical Informatics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Transplantation (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
US17/052,306 2018-05-03 2019-05-03 Oocyte-holding pipette for sperm injection methods without cytoplasmic aspiration Abandoned US20210230525A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
ESU201830624 2018-05-03
ES201830624U ES1214345Y (es) 2018-05-03 2018-05-03 Pipeta de sujecion de ovocitos para procedimientos de inyeccion espermatica sin aspiracion citoplasmatica
PCT/ES2019/070290 WO2019211503A1 (fr) 2018-05-03 2019-05-03 Pipette de maintien d'ovocytes utilisée dans des procédés d'injection spermatique sans aspiration cytoplasmique

Publications (1)

Publication Number Publication Date
US20210230525A1 true US20210230525A1 (en) 2021-07-29

Family

ID=62555864

Family Applications (1)

Application Number Title Priority Date Filing Date
US17/052,306 Abandoned US20210230525A1 (en) 2018-05-03 2019-05-03 Oocyte-holding pipette for sperm injection methods without cytoplasmic aspiration

Country Status (5)

Country Link
US (1) US20210230525A1 (fr)
EP (1) EP3777731B1 (fr)
CN (1) CN112367936A (fr)
ES (2) ES1214345Y (fr)
WO (1) WO2019211503A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3849706B1 (fr) * 2018-09-11 2023-06-14 Corning Incorporated Structure de pipette et procédés l'utilisant

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS63222680A (ja) * 1987-03-11 1988-09-16 Shimadzu Corp マイクロマニピユレ−シヨン用注射針装置
US5262128A (en) * 1989-10-23 1993-11-16 The United States Of America As Represented By The Department Of Health And Human Services Array-type multiple cell injector
US6048722A (en) * 1995-11-08 2000-04-11 Trustees Of Boston University Cellular physiology workstations for automated data acquisition and perfusion control
US6814936B1 (en) * 1999-07-01 2004-11-09 Goran Enhorning Pipette assembly having a small volume disposable tip
US20060228771A1 (en) * 1999-12-24 2006-10-12 John Dodgson Apparatus for and method of making electrical measurements on objects
US20070231879A1 (en) * 2006-03-30 2007-10-04 Steffen Hering Perfusion device for the treatment and examination of an object in a liquid
US20110250690A1 (en) * 2008-11-11 2011-10-13 Craig H Randall Microfluidic Embryo and Gamete Culture Systems
US20180291338A1 (en) * 2015-10-09 2018-10-11 Wolfgang Würfel Polar body injection

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1401342B1 (fr) * 2001-06-20 2005-01-12 Fertility Medical Equipment (Scotland) Limited Appareil et procedes destines a la preparation du sperme
CN2722861Y (zh) * 2004-07-07 2005-09-07 刘璋 微操作持卵器
CN101125105A (zh) * 2006-08-17 2008-02-20 吕祁峰 一种显微持卵管及其制作方法
WO2009088384A1 (fr) * 2008-01-07 2009-07-16 Willadsen Steen M Débit régulé de micropipettes
CN103249829B (zh) * 2010-08-20 2015-08-19 孙钰 用于自动化精子操作的系统和方法
DE102010040681A1 (de) * 2010-09-14 2012-03-15 Hamilton Bonaduz Ag Unterdruckgreifer zur Handhabung von Wirkstoff-Beads mit vorzugsweise konkaver Bead-Kontaktfläche
CN105838581B (zh) * 2016-05-11 2017-11-10 安徽农业大学 一种动物卵巢卵母细胞分离收集装置
ES1187335Y (es) * 2017-06-21 2017-10-02 Alcaide Francisco Vergara Pipeta de sujecion de ovocitos en procedimientos de inyeccion espermatica intracitoplasmatica
CN107649226A (zh) * 2017-09-21 2018-02-02 上海大学 一种微吸管及微吸管的制备方法

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS63222680A (ja) * 1987-03-11 1988-09-16 Shimadzu Corp マイクロマニピユレ−シヨン用注射針装置
US5262128A (en) * 1989-10-23 1993-11-16 The United States Of America As Represented By The Department Of Health And Human Services Array-type multiple cell injector
US6048722A (en) * 1995-11-08 2000-04-11 Trustees Of Boston University Cellular physiology workstations for automated data acquisition and perfusion control
US6814936B1 (en) * 1999-07-01 2004-11-09 Goran Enhorning Pipette assembly having a small volume disposable tip
US20060228771A1 (en) * 1999-12-24 2006-10-12 John Dodgson Apparatus for and method of making electrical measurements on objects
US20070231879A1 (en) * 2006-03-30 2007-10-04 Steffen Hering Perfusion device for the treatment and examination of an object in a liquid
US20110250690A1 (en) * 2008-11-11 2011-10-13 Craig H Randall Microfluidic Embryo and Gamete Culture Systems
US20180291338A1 (en) * 2015-10-09 2018-10-11 Wolfgang Würfel Polar body injection

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
English translation of JP-63222680 to Niwa et al, generated 2021 *
Otoi et al, Bovine Oocyte Diameter in Relation to Developmental Competence, Theriogenology, 48:769-774 (1997) *

Also Published As

Publication number Publication date
EP3777731A4 (fr) 2021-05-26
ES1214345U (es) 2018-06-19
EP3777731A1 (fr) 2021-02-17
EP3777731C0 (fr) 2023-11-01
WO2019211503A1 (fr) 2019-11-07
EP3777731B1 (fr) 2023-11-01
ES1214345Y (es) 2018-09-13
ES2964539T3 (es) 2024-04-08
CN112367936A (zh) 2021-02-12

Similar Documents

Publication Publication Date Title
Joris et al. Intracytoplasmic sperm injection: laboratory set-up and injection procedure
US20210230525A1 (en) Oocyte-holding pipette for sperm injection methods without cytoplasmic aspiration
US20140315282A1 (en) Microinjector mounting module and needle-holder component
EP3643578A1 (fr) Pipette de maintien d'ovocytes utilisée dans des procédés d'injection spermatique intracytoplasmique
CN107949335A (zh) 卵母细胞采集针
Pereira et al. Intracytoplasmic sperm injection: history, indications, technique, and safety
EP1401342B1 (fr) Appareil et procedes destines a la preparation du sperme
CN106479876B (zh) 一种连接器、转移装置及该转移装置的安装方法和使用方法
JP6862624B2 (ja) 卵細胞質内注入法
CN206930504U (zh) 一种用于去除离体血管内皮的灌注装置
JP2019154307A5 (fr)
CN202688311U (zh) 剥卵针
CN201067425Y (zh) 卵母细胞显微注射穿刺针
CN104323868A (zh) 基于附睾管腔内环境实验的微灌注方法及其装置
JP2020130181A (ja) インジェクションピペット
CN206157153U (zh) 一种新型多针取卵器
CN208435750U (zh) 显微操作持卵针
US10743974B2 (en) Follicular aspiration needle
Fernández et al. A new oocyte-holding pipette for intracytoplasmic sperm injection without cytoplasmic aspiration: an experimental study in mouse oocytes
CN209236324U (zh) 一种多功能血管鞘套件
CN208234922U (zh) 用于icsi的显微操作穿刺针
EP3166534B1 (fr) Dispositif d'introduction dans l'utérus
CN203988161U (zh) 脑室腹腔分流手术用皮下隧道构建系统及其通条
CN206391275U (zh) 防刺伤输液器
CN104083250B (zh) 一种视网膜下增殖条索支撑器

Legal Events

Date Code Title Description
STPP Information on status: patent application and granting procedure in general

Free format text: APPLICATION DISPATCHED FROM PREEXAM, NOT YET DOCKETED

STPP Information on status: patent application and granting procedure in general

Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION